JP2023509945A - Full-calorie, slow-digesting carbohydrate composition - Google Patents

Abstract

The present invention comprises at least 65% (w/w) of a glucose-based saccharide on a dry weight basis, comprising a reducing end and a D-glucose monomer linked by alternating α1-6 and α1-3 glycosidic linkages. and a maltose unit at the reducing end, comprising a glucose-based saccharide and 0.1 to 30% (w/w) fructose equivalent on a dry basis, wherein the glucose-based saccharide is A digestible carbohydrate composition having an average degree of polymerization greater than 12. [Selection drawing] Fig. 8

Description

[Background technology]
Carbohydrates have a unique role in the human diet. Carbohydrates serve technical functions such as texturizers or bulking agents. In some cases, carbohydrates can be considered sweeteners. Carbohydrates also provide energy and regulate blood sugar levels.

Ingestion of carbohydrates that are full-calorie, such as glucose syrup, maltodextrin, sugar, and starch hydrolysates typically result in high blood glucose peaks.

Current α-glucan components available on the market such as glucose syrups, isomalto-oligosaccharides, starch hydrolysates also compare their maximum concentrations (Cmax) and areas under the ascending curve (iAUC) to those of glucose. It is not very slow-digesting in that it is not sufficiently reduced by In addition, the unrefined α-glucan component, such as commercial scromalt, is high in monosaccharides, particularly fructose, and contributes to added sugars. Furthermore, raw starches such as wheat starch, which are known to have reduced Cmax and iAUC compared to glucose, are not sufficiently soluble and lose these properties upon heat treatment, especially in liquid applications.

Other α-glucan components of higher molecular weight such as indigestible dextrin, polydextrose, dextran and reuteran may not be full calorie because they are less digestible.

It is clear that there is a need to provide the market with an improved, full-calorie and slow-digesting carbohydrate composition that does not have the above drawbacks.

[Summary of Invention]
The inventors of the present application have surprisingly found a composition that does not have the drawbacks of prior art carbohydrate compositions. There is little contribution of added sugars, defined as containing no more than 5-10% sugars (monosaccharides and disaccharides). Little or no contribution of fructose. Furthermore, it is full calorie (3.5-4 kcal/g).

The compositions are also suitable for ready-to-drink (RTD) or ready-to-use (RTU) liquid matrices.

The composition is also suitable for ready-to-mix or powder applications.

In a first aspect, the present invention relates to a digestible carbohydrate composition, said digestible carbohydrate composition comprising:
a. At least 65% (w/w) of a glucose-based sugar on a dry weight basis having a reducing end and D-glucose monomers linked by alternating α1-6 and α1-3 glycosidic linkages a glucose-based sugar having an acceptor molecule at the reducing end; and
b. 0.1-30% (w/w) fructose equivalents on a dry basis,
The glucose-based sugars have an average degree of polymerization greater than 12 and the acceptor molecules are preferably maltose units.

In a second aspect, the invention relates to food products or beverages comprising said digestible carbohydrate composition.

In a third aspect, the invention relates to a method of reducing postprandial glucose in a subject comprising administering to a subject in need thereof an effective amount of said food product or beverage or said digestible carbohydrate composition. .

1 shows a scheme of a method according to the prior art; 1 shows a scheme of a method of making a digestible carbohydrate composition according to the invention. 1 shows a process design of a prior art method; 1 shows a process design of a method of manufacturing a digestible carbohydrate composition according to the invention. Fig. 3 is a chart showing the increase in molecular weight over conversion time when a constant supply of sucrose is applied; 1: fructose, 2: glucose, 3: DCC-1, 4: DCC-2 P1, 5: DCC-3 P2, 6: maltodextrin standards with different degrees of polymerization (DP). General research scheme. Postprandial glucose response after ingestion of DCC-1 and glucose syrup (25 g total carbohydrate). Increased postprandial glucose response after ingestion of DCC-1 and glucose syrup (25 g total carbohydrate). Postprandial glucose response after ingestion of DCC-2, DCC-3, and glucose syrup (33 g carbohydrate total). Increased postprandial glucose response after ingestion of DCC-2, DCC-3, and glucose syrup (33 g total carbohydrates). Mean relative iAUC of DCC-2 and DCC-3 compared to glucose syrup. Dark gray is the predicted relative iAUC when the product is pure glucose without fructose equivalents. Differences relative to the reference are significant (p<0.05) when the error bars do not exceed the 100% line. Postprandial breath hydrogen production after ingestion of DCC-2, DCC-3, and glucose syrup. Increased postprandial exhaled hydrogen production after ingestion of DCC-2, DCC-3, and glucose syrup.

Digestible Carbohydrate Compositions The present invention relates to a digestible carbohydrate composition, the digestible carbohydrate composition comprising at least 65% (w/w) glucose-based sugars on a dry basis, and reducing ends and , a D-glucose monomer linked by alternating α1-6 glycosidic bonds and α1-3 glycosidic bonds, and an acceptor molecule at the reducing end, a glucose-based sugar, and 30 % (w/w) of fructose equivalents, said glucose-based sugars having an average degree of polymerization greater than 12, and said acceptor molecules are preferably maltose units.

In particular, the present invention relates to a digestible carbohydrate composition comprising:
a. At least 65% (w/w) of a glucose-based sugar on a dry weight basis having a reducing end and D-glucose monomers linked by alternating α1-6 and α1-3 glycosidic linkages a glucose-based sugar having an acceptor molecule at the reducing end; and
b. 0.1-30% (w/w) fructose equivalents on a dry basis,
The glucose-based sugars have an average degree of polymerization greater than 12 and the acceptor molecules are preferably maltose units.

Acceptor molecules are preferably carbohydrates or carbohydrate derivatives.

Acceptor molecules may be selected from sugars or sugar alcohols that have free hydroxyl groups at one or more of carbon positions numbered 2, 3 and 6 that can accept glucose units from sucrose.

Carbohydrate acceptors are preferably sugars selected from the group consisting of maltose units, isomaltose units, maltitol units, (iso)maltotriose units and methyl-α-D-glucan units. Other preferred acceptor molecules are glucose units, gentiobiose units, raffinose units, melibiose units, isomaltitol units, isomaltooligosaccharide units, theanderose, kojibiose units, glucosyltrehalose units, cellobiose units, maltotetraose units, nigerose units, Lactose units, panose units, or mixtures thereof.

Preferably, the acceptor molecule is a maltose unit.

The average degree of polymerization was determined by GPC-RI (gel permeation chromatography with refractive index detection) or GPC-MALLS (gel permeation chromatography with multi-angle light scattering), or HPAEC-PAD (high performance using pulsed amperometric detection). anion exchange chromatography).

Preferably, the average degree of polymerization (or weight average degree of polymerization) is determined by HPAEC-PAD.

The average degree of polymerization can be 12-90, or 12-70, or 12-50, or 12-35, or 12-30, or 12-20, or 12-18 (HPAEC-PAD).

The average degree of polymerization can be greater than 13, 14, 15, or 16 (HPAEC-PAD). In some embodiments, the average degree of polymerization can be greater than 17 (HPAEC-PAD).

The average degree of polymerization can be less than 90, 70, 50, 35, 20, 19, or 18 (HPAEC-PAD).

The composition typically provides little contribution to added sugars in the form of mono- and disaccharides. Preferably, the composition contains 10% (w/w), 9% (w/w), 8% (w/w), 7% (w/w) added sugars in the form of monosaccharides and disaccharides. , 6% (w/w), or less than 5% (w/w).

In some embodiments, the composition has 65-99% (w/w) glucose-based sugars on a dry basis.

In some embodiments, the composition has 75-95% (w/w) glucose-based sugars on a dry basis.

In some embodiments, the composition has 85-90% (w/w) glucose-based sugars on a dry basis.

In some embodiments, the composition has at least 70% (w/w), preferably at least 75% (w/w) glucose-based sugars on a dry basis.

In some embodiments, the composition has at least 80% (w/w), preferably at least 85% (w/w) glucose-based sugars on a dry basis.

Compositions typically provide only a small amount of fructose equivalents on a dry basis.

In some embodiments, the composition comprises less than 25% (w/w) fructose equivalents on a dry basis.

In some embodiments, the composition comprises less than 20% (w/w) fructose equivalents on a dry basis.

In some embodiments, the composition comprises less than 10% (w/w) fructose equivalents on a dry basis. Preferably, the composition contains less than 9% (w/w), less than 8% (w/w), less than 7% (w/w), less than 6% (w/w), 5% (w/w), less than 3% (w/w), less than 2% (w/w), less than 1% (w/w), or less than 0.5% (w/w) fructose equivalents include.

In some embodiments, the composition comprises 0.1-25% (w/w), or 0.1-20% (w/w), or 0.1-15% (w/w) on a dry weight basis. /w), or 0.1-10% (w/w), or 0.1-9% (w/w), or 0.1-8% (w/w), or 0.1-7% (w/w), or 0.1-6% (w/w), or 0.1-5% (w/w), or 0.1-3% (w/w), or 0.1- 2% (w/w), or 0.1-1% (w/w), or 0.1-0.5% (w/w) of fructose equivalents.

In some embodiments, the fructose equivalent is leucrose, fructose, and/or sucrose. In some embodiments, the fructose equivalent is leucrose. In some embodiments, the fructose equivalent is fructose. In some embodiments, the fructose equivalent is sucrose. In some embodiments, the fructose equivalents are leucrose and fructose. In some embodiments, the fructose equivalents are leucrose and sucrose. In some embodiments, the fructose equivalents are fructose and sucrose. In some embodiments, the fructose equivalents are leucrose, fructose, and sucrose.

In one embodiment, the average molecular weight of the composition is greater than 2 kDa.

In one embodiment, the average molecular weight of the composition is 2-14.58 kDa.

The glucose-based sugars of the composition further comprise an α1-4 linkage at one terminus.

A typical composition of the invention comprises a glucose-based sugar having alternating alpha 1-6 and alpha 1-3 linkages, 1.5-2.5 grams of fructose equivalents per 33 grams of total carbohydrates, and 33 grams of total carbohydrates. 30.5-31.5 grams of glucose-based carbohydrate per gram, and the glucose-based sugar has an average degree of polymerization of 12-13.

Another exemplary composition of the invention comprises a glucose-based sugar having alternating alpha 1-6 and alpha 1-3 linkages, 2.5-3.5 grams of fructose equivalents per 33 grams of total carbohydrates, and 29.5-30.5 grams of glucose-based carbohydrates per 33 grams of carbohydrates, the glucose-based sugars having an average degree of polymerization of 17-18.

Typically the composition is full calorie. In some embodiments, the composition provides at least 3 kcal/gram, or 3.5 kcal/gram, or up to 4 kcal/gram, or 3.5-4 kcal/gram.

The composition induces a lower glycemic response in a subject compared to maltodextrin or glucose syrup. Typically, the glycemic response in subjects is lower in the three hour period immediately following ingestion of the composition. Glycemic response can be measured by area under the ascending curve (iAUC), eg, as described herein.

The composition has the added advantage of being slowly digested. Postprandial hydrogen production can be used as an indirect measure of digestibility. When indigestible carbohydrates reach the colon, they are fermented by colonic bacteria. This fermentation produces gases such as hydrogen and methane, which can be measured in the subject's exhaled breath.

In one embodiment, the composition does not provide more than 20 ppm hydrogen in breath in a period of 240 minutes immediately after ingestion of 33 grams of the composition by the subject. In one embodiment, 33 grams of the composition digests more slowly than 33 grams of maltodextrin syrup or glucose syrup.

The gastrointestinal tolerance of the composition is very high. In one embodiment, the composition does not cause one or more of diarrhea, abdominal cramps, vomiting, audible bowel sounds, or flatulence in a subject, for example, in a period of 180 minutes immediately after ingestion of 33 g of the composition. .

In some embodiments, the digestible carbohydrate composition according to the invention is for use in reducing the glycemic response in a subject, preferably a human subject.

In one embodiment, the composition is for use in reducing a glycemic response in a subject, the composition comprising a glucose-based saccharide having alternating alpha 1-6 and alpha 1-3 linkages; It contains 2.5-3.5 grams of fructose equivalents per 33 grams of total carbohydrates, 29.5-30.5 grams of glucose-based carbohydrates per 33 grams of total carbohydrates, and has an average DP of 17-18.

In one embodiment, the composition is used to reduce a subject's glycemic response during a period of 60, 120, or 180 minutes immediately after ingestion of the composition as compared to the glycemic response to glucose syrup in the subject. It is for

Glycemic responses can be reduced by up to 45% in subjects during the 60 minute period immediately after ingestion. Glycemic responses can be reduced by up to 38% in subjects during the 120 minute period immediately following ingestion. Glycemic responses can be reduced by up to 33% in subjects during the 180 minute period immediately following ingestion.

Preferably, the subject is a human subject.

Food Products or Beverages The present invention also relates to food products or beverages comprising the digestible carbohydrate compositions described herein. The digestible carbohydrate composition may be in the form of a powder, such as a ready-to-mix powder for beverages. In other embodiments, the digestible carbohydrate composition may be in the form of a liquid, eg syrup.

Preferably, the beverage is a ready-to-drink (RTD) or thermally treated beverage.

In some embodiments the food product or beverage is a dietary supplement.

In some embodiments the food product or beverage is a nutritional product.

The nutritional formulation can be, for example, any oral nutritional form such as health drinks, commercial beverages, optionally juices, milkshakes, yogurt drinks, smoothies or soft drinks including soy-based beverages, nutrition bars, or any types of foods such as baked goods, cereal bars, dairy bars, snack foods, soups, breakfast cereals, muesli, candy, tablets, cookies, biscuits, crackers (such as rice crackers), and dairy products may have been

A nutritional formulation may be a nutritional bar.

A nutritional formula may further include fat, protein, and other carbohydrate sources.

Dietary supplements or formulations can be, for example, tablets, capsules, lozenges, or liquids. Dietary supplements or formulations may contain protective hydrocolloids (gums, proteins, modified starches, etc.), binders, film formers, encapsulants/materials, barrier/shell materials, matrix compounds, coatings, emulsifiers, surfactants, Solubilizers (oils, fats, waxes, lecithins, etc.), adsorbents, carriers, fillers, co-compounds, dispersants, wetting agents, processing aids (solvents), flow agents, flavoring agents, bulking agents Agents, jelling agents, and gel forming agents may also be included.

In some embodiments, the nutritional product or dietary supplement is for use as a medicament.

In some embodiments, the nutritional product or supplement is for managing glycemic control or reducing glycemic response in a human subject, eg, a healthy human subject.

In some embodiments, the nutritional formulation or dietary supplement is for an infant, child, or adolescent human subject.

In some embodiments, the nutritional product or dietary supplement is for a diabetic and/or pre-diabetic human subject.

In some embodiments, the nutritional product or supplement is suitable for human subjects under acute care.

In some embodiments, the nutritional formulation or dietary supplement is suitable for weight loss products for human subjects.

In some embodiments, the food product is a pet food product.

Methods of Reducing Postprandial Glucose The present invention also provides for administering an effective amount of a digestible carbohydrate composition as described herein or a food product or beverage as described herein to a subject in need thereof. methods of reducing postprandial glucose in a subject, comprising:

In some embodiments, the subject is a human subject.

In some embodiments, the subject is a companion animal subject, such as a dog or cat.

DEFINITIONS All percentages given herein are by dry weight by total dry weight of the composition, unless otherwise indicated. As used herein, “about,” “approximately,” and “substantially” refer to values within a numerical range, such as −10% to +10% of the reference number, preferably −5 of the reference number. % to +5%, more preferably −1% to +1% of the reference number, most preferably −0.1% to +0.1% of the reference number. understood. All numerical ranges herein should be understood to include all integers or fractions within the range. Moreover, these numerical ranges should be interpreted to support claims that are directed to any number or subset of numbers within the range. For example, a disclosure of 1-10 should be interpreted to support ranges of 1-8, 3-7, 1-9, 3.6-4.6, 3.5-9.9, and so on.

As used in this disclosure and the appended claims, the singular forms "a," i.e., "a," "an," and "the," include plural references unless otherwise indicated. be Thus, for example, reference to "a component" or "the component" includes two or more components.

The terms "comprise," "comprises," and "comprising" are to be interpreted as inclusive rather than exclusive. should. Similarly, the terms "include," "including," and "or" are all to be construed as inclusive unless the context clearly dictates such an interpretation. is. However, the compositions disclosed herein may not contain elements not specifically disclosed herein. Thus, the disclosure of embodiments using the term "comprising" includes both embodiments "consisting essentially of" the identified components, and embodiments "consisting of ” includes disclosure of embodiments.

Digestible carbohydrate composition: A carbohydrate composition that is digested in the small intestine and does not reach the colon does not result in exhaled hydrogen production upon ingestion by a subject, preferably a human subject.

Carbohydrate Composition: All carbohydrates including digestible carbohydrates and sugars.

Glucose-based sugars: Carbohydrates of various sizes and molecular weights (MW) containing glucose monomers as building blocks linked by α-glycosidic bonds.

Maltose unit: A disaccharide composed of two glucose units linked by an α1-4 glycosidic bond.

Reducing end: The reducing end of a carbohydrate is a monosaccharide with a free anomeric carbon that does not participate in glycosidic linkages and thus can be converted to the open chain form.

Fructose Equivalents: Any carbohydrate, including monosaccharides and disaccharides, is composed of at least one fructose monomer building block and is subject to fructose metabolism in the small intestine upon ingestion by a subject, preferably a human subject. Examples are leucrose, sucrose and fructose.

Average Degree of Polymerization (DP): The average number of monosaccharide building blocks per glucose-based sugar chain in each carbohydrate composition.

Maltose-alternan-oligosaccharide (MAOS): An example of a glucose-based sugar with maltose at the reducing end.

Monosaccharides: Any class of simple sugars that can serve as building blocks of larger carbohydrate structures. Monosaccharides cannot be hydrolyzed to yield simpler sugars. Examples of monosaccharides include glucose and fructose. Disaccharide: Any sugar containing two constituent monosaccharides, such as sucrose, leucrose, and maltose.

Added Sugars: All the above monosaccharides and disaccharides present in the composition Glycemic Response or Postprandial Glucose: Glucose concentration measured in blood or interstitial tissue after ingestion of carbohydrate by a subject, preferably a human subject.

The terms "food", "food product", and "food composition" mean a product or composition intended for consumption by an individual, such as a human, to provide such individual with at least one nutrient. As used herein, these terms encompass any form of food, including both liquids (eg, beverages) and solids. The compositions of the present disclosure, including many of the embodiments described herein, are useful in the elements disclosed herein as well as in diets whether or not described herein. may comprise, consist of, or essentially comprise any additional or optional raw materials, components or elements that are

A "beverage" is a substantially homogeneous liquid that is at least 85% water, in some embodiments at least 90% water or at least 95% water by weight. A "ready-to-drink" beverage is in liquid form that can be consumed without the addition of further liquid and is preferably sterile. Reconstitution and dilution can comprise adding water and/or milk to the powder or concentrate, respectively, and in some embodiments the method comprises a reconstitution or dilution step.

Those skilled in the art will understand that they can freely combine all the features of the invention disclosed herein. In particular, features described for the product of the invention may be combined with the method of the invention, and vice versa. Furthermore, features described for different embodiments of the invention may be combined. Where known equivalents exist for particular features, such equivalents may be incorporated as specifically noted herein.

Further advantages and features of the invention are evident from the figures and the non-limiting examples.

Example 1
Production of Digestible Carbohydrate Composition Prior art methods are known, for example from WO 0047727 (A2) and WO 2009095278 (A2), comprising steps P1 to P4 of FIG. 1a .
In step P1, bioconversion of sucrose and maltose is carried out in a batch reactor at T=37° C. for about 20 hours. The sucrose:maltose ratio is chosen to be 7:1 (w/w or mol/mol). Step P1 is carried out in a bioconversion batch reactor shown in FIG. 2a.

P2 is a step of ultrafiltration to remove alternan polymer (alternan polysaccharide) and alternansucrase enzyme (AlSu). This step takes place in the ultrafiltration device shown in Figure 2a.

In P3 fructose is removed by nanofiltration. This step takes place in the nanofiltration device shown in Figure 2a. Here water is added and the mixture of water and fructose is removed.

In the final step P4, the product from P3 is concentrated by evaporation. This step takes place in the evaporator of FIG. 2a and yields maltose-alternan-oligosaccharides (MAOS).

The method of the invention comprises, in a particular embodiment, three steps S1-S3 shown in FIG. 1b.

Compared to the prior art process of FIG. 1a, the bioconversion in step S1 involves continuous supply of sucrose and continuous removal of fructose. Semi-continuous supply and removal is possible. The mass ratio of sucrose (total added) to maltose is 19:1 (19 kg sucrose/1 kg maltose) and the duration is about 72 hours. Since the fructose is already removed in step S1, the prior art step P3 can be omitted. By removing fructose, a higher sucrose to maltose ratio can be used and a higher degree of polymerization of maltose-alternan-oligosaccharides can be reached.

Step S1, bioconversion takes place in the reactor of Figure 2b, maltose and alternan sucrase enzyme (AlSu) are present in water. As described above, the biotransformation consisted of continuous supply of sucrose, continuous removal of fructose, a duration of about 72 hours (variable), T=37° C., and a sucrose:maltose ratio (w/w or mol/ mol). Sucrose is added as feed (dissolved in water) and the contents of the reactor are agitated. In the biotransformation in the reactor, alternan-oligosaccharides containing the acceptor molecule maltose (also called maltose-alternane-oligosaccharides (MAOS)) are formed as major products, and alternan polymers, fructose, and leucrose are by-products. formed as The contents are continuously circulated from the reactor through a membrane cell (diafiltration cell) and water, fructose and leucrose are removed by membrane filtration. In this example, membrane filtration is nanofiltration performed as constant volume diafiltration. The leucrose content is reduced to less than about 30% to 10% compared to the prior art. The removed water is replaced by the water feed stream.

The reactor and membrane cell form a combined bioconversion and nanofiltration device, also called a reactor system.

Method step S2 of this embodiment corresponds to step P2 of the prior art. Here, alternan polysaccharide (alternan polymer) and alternansucrase enzyme (AlSu) as by-products are removed by ultrafiltration. This step is beneficial if desired if more alternan polymer is formed or to lead to DPw of the desired alternan species remaining.

Method step S3 of this embodiment corresponds to step P4 of the prior art. In this step the product is concentrated by evaporation.

Table 1 shows the composition of the reaction solution used during the production. The 2.1 L of solution totals about 5.6 L of water in the system (dead volume).

The method is run without depletion for the first hour to minimize the risk of depletion of maltose across the membrane. Thereafter fructose is constantly depleted by nanofiltration membranes Filmtec NF270-2540 (DOW). Once chain extension was completed, the nanofiltration module was replaced with a TRISEP2540-UE50-QXF ultrafiltration module (Microdyn Nadir). This separated the maltose-alternan-oligosaccharide (MAOS) fraction from the longer aged chains and enzymes. The membranes used in this method and the method parameters used are summarized in Table 2. The filtrate was finally concentrated to a dry matter content of >72%.

Figure 3 shows the increase in chain length over the course of the method. Values were recorded by GPC-RI measurement. The relationships for calculating DPw from the Mw values in FIG. 3 are as follows. DPw=Mw/(162 Da). It can therefore be seen that at the end of the method a DPw of about 15.4 is reached (2500/162).

To reach an average chain length DPw of about 15, 19 kg of sucrose (total) was used per kg of maltose.

Comparative Example: Alternan-oligosaccharides were produced according to the method shown in Figure 1a. A 21:1 ratio (kg sucrose:kg maltose) was used resulting in a DPw of 9.9. For the measurement of DPw, GPC-RI was used, but not exactly the method-protocol described above. Nevertheless, compared to the results in Figure 3, the present invention shows that alternan with higher DPw is obtained.

Table 4 summarizes the DPw values of two samples obtained by the method of the invention analyzed by the HPAEC-PAD method.

Example 2
Structural Analysis of Digestible Carbohydrate Compositions Prepared According to Example 1 Monosaccharides (glucose, fructose, leucrose, sucrose, maltose) were subjected to Quantification was performed using an inert styrene divinylbenzene polymer (Dionex Corporation, 2010) and a Dionex ICS-3000 DC instrument equipped with gold triple potential pulsed amperometric detection (PAD). Eluent A (300 mM aqueous NaOH), eluent B (MiliQ water), and eluent C (500 mM CH3COONa in 150 mM NaOH) were used as mobile phases in gradient mode with a total run time of 35 minutes.

The monosaccharides present on a dry basis in the test product are summarized in Table 3 below.

DPw values were measured by HPAEC-PAD after reduction and hydrolysis of glucose-based sugars. Two milliliters of solution containing 6 g/mL digestible carbohydrate composition was treated with 0.2 mL of NaBH4 solution (40 mg/mL) in 0.5 M ammonia at 40°C for 30 minutes. The reduced sample was then hydrolyzed with 0.5 mL of 2M trifluoroacetic acid and heated at 121° C. for 1 hour to release the monomer. The released monomers were analyzed by injecting the sample solution into a Thermo Scientific™ Dionex™ ICS-6000 ion chromatograph system equipped with a CarboPac™ MA1 and eluting with eluent (water and NaOH 1000 mM) at 0.4 mL/min. was quantified by feeding The DP value is calculated by the following formula.

The DP values of the samples are summarized in Table 4 below.

The molecular weights of DCC-1, DCC-2 P1, and DCC-3 P2 were also determined using silica TLC plates, a mixture of chloroform:acetic acid:water:ethanol (30:35:25:20) as mobile phase and diphenylamine- Quantitative analysis was performed using thin layer chromatography using an aniline reagent (Fig. 4).

Glycosidic binding profiles of glucose-based oligosaccharides were determined on partially methylated alditol acetates by GC-MS. Briefly, samples were dissolved in anhydrous DMSO, deprotonated by the addition of n-butyl lithium (Sigma 230707) and methylated with methyl iodide (Sigma 289566). Methylated samples were then hydrolyzed with 2N TFA (60 min at 121°C). Hydrolyzed samples were evaporated under a stream of nitrogen, redissolved in 1 M ammonium hydroxide, and aldehyde groups were reduced with a DMSO solution containing sodium borate (20 mg/ml). Glacial acetic acid was added dropwise to quench the reaction, and 1-methylimidazole and acetic anhydride were added to effect acetylation. Partially methylated alditol acetates in acetone were purified by GCM (7890A-5975C MSD, Agilent Technologies, Inc., Santa Clara, Calif., USA) on a Supelco 24111-U SP-2380 capillary column (0.5 μL injector volume, injector temperature 250° C., detector temperature 250° C., carrier gas, helium: 30 mL/min, split ratio 40:1, temperature program: 100° C. for 3 min, 4° C./min to 270° C. for 20 min). Electron impact spectra were acquired at 69.9 eV over the mass range of 50-550 Da.

Higher values of 1,6-glycosidic linkages contribute to the amount of 1,5,6-tri-O-acetyl-1-deuterio-2,3,4-tri-O-methyl-D-glucitol observed. is explained by the leucrose content in the digestible carbohydrate composition. In addition, leucrose and monomeric glucose contribute to the amount of terminal Glc in the digestible carbohydrate composition.

Example 3
Preparation of Digestible Carbohydrate Compositions In this example, the parameters of the process described in Example 1 for DCC-3 were modified as shown in the table below, and the alternan sucrase was divided into four equal parts for the reaction. A first portion was present before the sucrose was fed to the reactor.

Enzymatic activity refers to the total units of alternansucrase enzyme used in the method.

The reaction time of the process of the invention (not including further process steps such as removal of alternan polysaccharide and alternan sucrase enzyme by further membrane filtration or concentration of the retentate obtained in further membrane filtration) is determined by the amount of sucrose can be shortened by increasing , increasing sucrose feed rate, increasing enzyme activity, and increasing temperature.

Glycosidic binding profiles of glucose-based oligosaccharides were determined according to Example 2. The results are shown in Table 6 below.

Example 4
Results of Preliminary Crossover Studies In a preliminary randomized controlled crossover study, a digestible carbohydrate composition with a degree of polymerization of 7 (DCC-1) was tested in 16 healthy volunteers. 25 g of DCC-1 dissolved in 300 mL of water was ingested and the postprandial glucose response was measured at 2 hours (Figures 6 and 7). As a control, 25g of glucose syrup was ingested.

In this study, ingestion of DCC-1 resulted in lower glycemic responses compared to glucose syrup, but there were no significant differences for tmax, iCmax, and iAUC. Additionally, no severe gastrointestinal discomfort was reported by study participants. These results indicate that ingested α-glucan is largely digested but does not provide a significant benefit on glycemic response due to a small increase in degree of polymerization (DP7) compared to glucose syrup. suggesting.

Example 5
CLINICAL STUDY - METHODS Monocentric, controlled, randomized, double-blind, crossover with 16 participants ingesting different test products containing different types of carbohydrates with different structure and composition. did the test. A subject's participation in this research project is voluntary and may be terminated at any time without the need to withdraw consent or justify termination of participation. If a participant declined, the data collected and coded up to that point was used and anonymized after analysis.

The purpose of this study was to examine the effects of different α-glucans on postprandial glucose response (PPGR) and hydrogen (H2) production to provide insight into their digestibility.

The first objective was to determine if the α-glucans tested induce lower glucose responses compared to maltodextrin.

A secondary objective was to indirectly determine whether the α-glucans tested were completely absorbed in the small intestine and induced gastrointestinal discomfort.

Glucose responses were assessed during 3 hours after ingestion of the test product. The 3-hour area under the ascending curve (3h-iAUC) was the primary endpoint to address the primary objective.

As secondary endpoints, the following parameters were analyzed:
Other parameters derived from PPGR (i.e. cross-sectional values, iCmax, Cmax, tmax, partial iAUC, AUC);
Digestibility by measuring exhaled H2 for 4 hours after ingestion of test product. From these H2 curves, the parameters of 4h-iAUC, partial iAUC, partial AUC, Cmax, and cross-sectional values were derived. All these values were derived from H2 data corrected for CO2. Similar parameters were derived from H2 data uncorrected for CO2 and CH4 data (with or without CO2 correction);
Gastrointestinal tolerance 3 hours after ingestion of product by visual analogue scale for each symptom of interest: 1) diarrhea, 2) abdominal cramps, 3) vomiting, 4) audible bowel sounds, 5) flatulence or gas.

Glycemia was measured using a flash glucose monitoring (FGM, Free Style Libre®, Abbott) device for minimally invasive, real-time monitoring of blood glucose levels in interstitial fluid. . FGM has been developed and validated for use in adults with type 1 or type 2 diabetes [1-4] and measures interstitial glucose every 15 minutes for 14 days.

The breath hydrogen test is widely used to detect carbohydrate malabsorption. Its principle relies on the detection of hydrogen in exhaled breath primarily due to bacterial fermentation of carbohydrates in the colon. In order to obtain rapid and reliable indirect information on carbohydrate absorption, this study used a breath analyzer (Lactotest 202, M.E.C Belgium) measuring H2.

The test products were α-glucans with varying degrees of polymerization (DP) and glycemic binding, and thus different degrees of digestibility. Digestible Carbohydrate Composition 2 (DCC-2) has a DP of 12.4 and Digestible Carbohydrate Composition 3 (DCC-3) has a DP of 17.3. As a reference, fully digestible and full-calorie maltodextrin (glucose syrup) was administered. The test product contained 33 g of total carbohydrate in either powder (glucose syrup) or syrup (digestible carbohydrate composition) form. The day before the test visit, the test product was dissolved in 300 mL of water and stored overnight at 4°C. The morning before consumption, the beverages were warmed to room temperature before being served to participants.

The subject population consisted of fully healthy men and women aged 18-45 years (based on medical history) with a BMI of 20-29.9 kg/m2. Subjects were ineligible to participate if they presented one or more of the following exclusion criteria:
women who are pregnant or breastfeeding;
Four weeks prior to the study, concomitant medications that may affect study procedures and assessments, such as antibiotics, antacids, or other medications that affect transit time, colonoscopy, enema, or other bowel cleansing procedures;
Serious medical/surgical events within the past 3 months that may compromise study procedures and evaluations;
Abnormal intestinal transit and history of chronic constipation with spontaneous bowel movements on average <3 times per week or chronic or recurrent diarrhea with spontaneous bowel movements >3 times per day;
known food allergies and intolerances to the test product;
medically known skin sensitivities to adhesives and plasters;
Alcohol consumption in excess of 2 drinks per day. 1 glass is 0.4 dl for strong alcohol, 1 dl for red or white wine, or 3 dl for beer;
smoker;
Volunteers who cannot be expected to comply with the protocol;
A subject who has a hierarchical relationship with a member of the study team.

An announcement was made within Nestle entities and potential candidates were contacted. Volunteers who wished to participate and who were eligible received a full oral briefing (V0) by the study team regarding the purpose, methods and possible risks/disadvantages of the study. Candidates were invited to a medical screening visit to confirm eligibility and sign informed consent.

One FGM device was worn on each volunteer's non-dominant arm at least 24 hours prior to the first test product intake. Participants then attended different test visits for each of the beverages at least one day apart so that they could consume a low-fiber diet the day before each test visit. After completion of all measurements, the sensor was removed on the final day of testing.

Subjects were required to abstain from alcohol the day before each study visit. Medications such as aspirin or dietary supplements containing vitamin C, stimulants, laxatives and antibiotics that could affect FGM measurements were asked to be avoided 4 weeks prior to and during the entire study. Fermentable foods such as complex carbohydrates should be avoided the day before the breath test. The final meal on the day before the test had to be fiber-free and not overly satiating. Participants were also asked to avoid chewing gum from 8:00 pm the day before each study visit.

A general study scheme is described in FIG. Subjects arrived at the Metabolic Department at 8:00 pm after fasting from 8:00 am the previous day. To minimize interference with breath testing, subjects should avoid chewing gum, avoid the use of perfume, and be asked to brush their teeth thoroughly at home before each visit in the metabolic department. was taken.

Readings from the FGM device were taken before and after ingestion of the product. The average of the two measurements was taken as the baseline value. Readings were also taken at 30, 60, 90, 120, 150, 180, 210 and 240 minutes. Breath samples for determination of hydrogen production were obtained prior to product ingestion and then immediately after glucose reading at 30, 60, 90, 120, 150, 180, 210, and 240 minutes. . After 180 minutes of product ingestion and upon completion of the corresponding glucose readings and breath tests, subjects were asked to complete a questionnaire regarding gastrointestinal symptoms. After these procedures, a fiber-free breakfast was provided consisting of white bread, honey, and coffee or tea or water. Any other food, including chewing gum, was prohibited during the study and only water was allowed.

A minimum sample size of N=10 subjects completing the test is required by ISO-26642 for determining the glycemic index (GI) of food products and classifying them [5]. For α-glucan, test 17.08. BIO used N=16 to detect a 35% reduction in PPGR 3h-iAUC compared to the maltodextrin control (SD=50%) with α=5% (two-tailed) and power=80%. is required. Because this preliminary study used smaller serving sizes (i.e., 25 g), the calculated sample size is conservative (i.e., variability in healthy subjects is generally reduced with larger serving sizes). be done). This sample size is also suitable for H2-related endpoints, as indicated by the fact that N=16 allows a difference of 0.35 log ppm to be distinguished with α=5%, power=80%. [6]. In conclusion, the sample size for this study was N=16.

For the primary endpoint (ie, PPGR-derived 3h-iAUC), the test product was compared to the maltodextrin control using a paired t-test (according to ISO-26642 logic). The Benjamini-Hochberg procedure was applied to control the false discovery rate (FDR) at α=5% [7]. Sensitivity analyzes were performed using mixed models to take into account potential systematic position or carryover effects [8]. For all other pairwise comparisons and all other endpoints the analyzes were the same, but were not corrected for multiplicity.

Example 6
Clinical Trial Results Healthy subjects with a mean age of 31.4±5.9 years, a mean BMI of 23.0±1.6 kg/m2, and a mean fasting blood glucose level of 4.8±0.5 mmol/L. Sixteen volunteers (6 females and 10 males) were recruited.

Figures 8 and 9 show the 4-hour postprandial glucose response and incremental glucose response, respectively, for all test products. Compared to glucose syrup, both glucose-based sugars resulted in significantly lower iCmax, but no significant difference is observed in tmax. The time required for blood glucose levels to return to baseline values is not significantly longer compared to the reference. 1h-iAUC was significantly lower for both test products, but only DCC-3 resulted in significantly lower 2h- and 3h-iAUC compared to glucose syrup.

Comparing the glucose curve shapes of the α-glucans with the reference, lower glucose peaks and slower return to baseline are observed for both α-glucans. The drop in blood glucose after the peak was slower than in the reference, with blood glucose remaining slightly elevated for a longer period of time, resulting in α-glucan being digested and utilized slightly more slowly than maltodextrin. It suggests that Furthermore, α-glucan intake results in lower hypoglycemia after returning to baseline compared to controls.

FIG. 10 represents the mean relative 1h-, 2h- and 3h-iAUC of the test product compared to glucose syrup. DCC-2 causes a 27% (p<0.05), 16% and 13% reduction in 1h-, 2h- and 3h-iAUC, respectively, compared to reference. DCC-3, which has a higher degree of polymerization than DCC-2, produced greater hypoglycemia compared to the reference, 45% (p<0.05) for 1h-iAUC and 38% for 2h-iAUC (p<0.05). p<0.05) and 33% for 3h-iAUC (p<0.05).

DCC-2 and DCC-3 contain 2.1 and 2.9 g of fructose equivalents respectively and are therefore not fully glucose-based, unlike glucose syrups. The dark gray bars in FIG. 10 are the predicted relative iAUC corrected data when the product is a pure glucose system. After correction, the reduction of iAUC for DCC-2 remains significant only for 1h-iAUC (24%), whereas the reduction of DCC-3 1h-, 2h- and 3h-iAUC is significant (42% each). %, 34% and 28%). The reduction in blood glucose is significant even with the corrected predictions, and it can be concluded that fructose is not the main factor in the PPGR reduction.

Postprandial exhaled H2 was used to indirectly measure product digestibility. When indigestible carbohydrates reach the colon, they are fermented by colonic bacteria. This fermentation produces gases such as H2 and CH4, which can be measured in the subject's exhaled breath. After ingestion of the test products no significant difference was observed from each other and compared to the fully digestible control product (Figures 11 and 12). A typical range for fasting breath hydrogen is 7±3 ppm, and the 240 minute maximum is found within this range. Only DCC-2 had higher values than the other products, but remained below the carbohydrate malabsorption threshold of 20 ppm and decreased during the study to reach levels similar to the other products. At 180 minutes after eating breakfast, a slight increase in hydrogen production was observed, which Lactotest suggests may result from the presence of very small amounts of complex carbohydrates or slightly incomplete absorption of simple carbohydrates (e.g., fructose). , suggesting the detection of even smaller amounts of H2. Therefore, all α-glucans appear to be highly digestible and not induce fermentation of carbohydrates such as fiber.

Gastrointestinal tolerance to the product was assessed using a visual analogue scale for 5 different symptoms: cramping, bowel sounds, diarrhea, flatulence and vomiting (Table 6). In general, few people reported discomfort and no severe events were reported. In fact, bowel sounds were the symptom with the highest score. Scores from those reporting discomfort were mostly low. The average z-score remains low and is the highest for the reference product glucose syrup. Thus, in healthy subjects gastrointestinal tolerance to α-glucan is good, suggesting that it is largely digestible.

As expected, all α-glucans tested produced lower postprandial glycemic responses compared to the fully digestible maltodextrin control. α-Glucan structures with alternating α1-3/6 linkages lead to a significant reduction in glucose response compared to glucose syrup when the molecular weight is higher than 1.6 kDa (DP>10). Indeed, in healthy volunteers, the glucose-based saccharide DP17.3 gave a 38% (p<0.01) reduction in glucose response (2h-iAUC).

Unlike fiber, which undergoes colonic fermentation due to incomplete digestion, the results of this clinical trial indicated that none of the products tested produced exhaled hydrogen as a result of colonic fermentation, suggesting that α- suggesting that the glucans are mostly digestible. Furthermore, the gastrointestinal tolerance assessment did not differ from that reported for the reference product, indicating very low discomfort.

References 1. Distiller, L.; A. , I. Cranston, andR. Mazze, First Clinical Experience with Retrospective Flash Glucose Monitoring (FGM) Analysis in South Africa: Characterizing Glycemic Control with Ambulatory Glucose Profil. J Diabetes Sci Technol, 2016.10(6): p. 1294-1302.
2. Bonora, B.; , et al. , Head-to-head comparison between flash and continuous glucose monitoring systems in outpatients with type 1 diabetes. J Endocrinol Invest, 2016.39(12): p. 1391-1399.
3. Schierenbeck, F.; , A. Franco-Cereceda, andJ. Liska, Accuracy of 2 Different Continuous Glucose Monitoring Systems in Patients Undergoing Cardiac Surgery. J Diabetes Sci Technol, 2017.11(1):p. 108-116.
4. Akintola, A.; A. , et al. , Accuracy of Continuous Glucose Monitoring Measurements in Normo-Glycemic Individuals. PLoS One, 2015.10(10): p. e 0139973.
5. Internal Standard Organization, Food Products-Determination of the Glycaemic Index (GI) and Recommendations for Food Classification. ISO 26642.2010.
6. Grysman, A.; , T. Carlson, and T. M. Wolever, Effects of sucromalt on postprandial responses in human subjects. Eur J Clin Nutr, 2008.62(12):p. 1364-71.
7. Benjamini, Y.; and Y. Hochberg, Controlling the False Discovery Rate: a Practical and Powerful Approach to Multiple Testing. J Roy Stat Soc, 1995.57:p. 289-300.
8. Senn, S.; , Cross-over Trials in Clinical Research. 2002, Chichester: New York: J.M. Wiley.

Claims (16)

a. At least 65% (w/w) of a glucose-based sugar on a dry weight basis having a reducing end and D-glucose monomers linked by alternating α1-6 and α1-3 glycosidic linkages a glucose-based saccharide, wherein an acceptor molecule is present at the reducing end;
b. 0.1-30% (w/w) fructose equivalents on a dry basis,
A digestible carbohydrate composition, wherein said glucose-based sugars have an average degree of polymerization greater than 12 as measured by HPAEC-PAD, and said acceptor molecules are preferably maltose units. 2. The digestible carbohydrate composition of claim 1, wherein said average degree of polymerization is greater than 17 as measured by HPAEC-PAD. 3. A digestible carbohydrate composition according to claims 1 and 2, wherein said composition has at least 80% (w/w), preferably at least 85% (w/w) glucose-based sugars on a dry basis. The composition has α1-6 branch points on glucose 3-linked by said alternating α1-6 glycosidic linkages and α1-3 glycosidic linkages in greater than 0.1% (w/w) of total glycosidic linkages. A digestible carbohydrate composition according to claims 1-3. A digestible carbohydrate composition according to claims 1-4, wherein said composition comprises 0.1-10% (w/w) fructose equivalents on a dry basis. 6. A digestible carbohydrate composition according to claim 5, wherein said fructose equivalent is leucrose, fructose and/or sucrose. A digestible carbohydrate composition according to claims 1-6 for use in reducing the glycemic response in a subject. A food product or beverage comprising a digestible carbohydrate composition according to claims 1-6. 9. The food product or beverage of claim 8, wherein said food product or beverage is a dietary supplement. 10. Food product or beverage according to claims 8 and 9, wherein said food product or beverage is a nutritional formulation. 11. The food product or beverage of claim 10, wherein the nutritional product is for a) diabetic and/or pre-diabetic human subjects, or b) infant, child or adolescent human subjects. 12. The food product or beverage according to claims 10 and 11, wherein said nutritional product is for human subjects under acute care. The food product of claims 8-10, wherein the food product is a pet food product. reducing postprandial glucose in a subject, comprising administering an effective amount of a digestible carbohydrate composition according to claims 1-6 or a food product or beverage according to claims 8-13 to a subject in need thereof. How to reduce. 15. The method of claim 14, wherein said subject is a human subject. 15. The method of claim 14, wherein said subject is a companion animal subject.

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